TdT -Summary

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From:Daniel & Linda Botsford <> (by way of histonet)
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Thanks to all who responded.
1.DAKO carries a polyclonal TdT (A3524) that requires high pH, citrate
heat target retrieval.
2. Dako's TdT catalog# A3524  at a 1:10 dilution with a thymus as a
control.  I also use their LSAB2 kit as a detection system but I'm sure
other detection systems will work too.  I pretreat with a citra
step for 15 minutes. I'm sure that other pretreatments will work also.
3.We use the antibody from SuperTech at a 1:10 dilution with the
decloaker pressure cooker for 20 min. in citrate buffer ph 6 hier
the detection is a vector quick kit LSAB hrp system.
4. We use DAKO products,...We use high pH heat induced epitope
retrieval. Then incubate in primary TdT antibody @1:10 dilution for 30
minutes (room temp), and 30 minutes secondary and tertiary. The results
aren't the best I've seen for this antibody and we're still trying to
"tweak it", but it is readable and our doctors don't grumble (too much).

5.We use Supertech's pAb to Tdt at 1:200 (overnight primary antibody
incubation) with
DAKO's High pH retrieval solution (30' steam heat).  Zymed also has a
pAb that works well
in formalin-fixed, paraffin-embedded tissue.
6. Steam retrievel using Dako high pH Target Retrievel buffer - 20 min.
we used the antibody from Serotec cat# 004, rabbit anti-Tdt (18003270286
) titre at 1:10 & 1:20 using normal thymus as a control.
Note: our lab ran both titres on each case as 1:30 did not produce good
results at all. It was either one or the other but never consistantly
either of them.
7.I use Intergen Apotag with a Proteinase K pretreatment. Work well in
8.I have worked with zymed and other manufacturers Tdt antibody and have
control slides made for a method done in our hematology dept. I use
retrieval and incubate for 32 min and have done this on both the ES and
the Nexes.I got
great results.
9.We use the TdT from Dako (A3524) at a 1:20 dilution overnight
Microwave pretreatment in Citric acid buffer pH 6.0. Detection either
the StreptABC/HRP Duet kit/ Envision works well.
10.InnoGenex carries 4 TdT antibodies(AM2465,AM2466,AP2467,AP2468). All
4 are
validated for paraffin sections. All four are diluted 1:100 for IHC. The

data sheets, which include pre-treatments, can be accessed online at:
11.We do Tdts on our NexES using the Super Tech antibody at a 1:5
dilution for 32 minutes.We have found this dilution to be stable for
about 2 weeks.  We use an EDTA buffer retrieval (steam) and an A/B
block.....tissue should be cut fresh - only stable about 2 weeks also.We
have gotten good staining, but because it is a poly there is that
"blush" that will show up -especially on megakaryocytes - although it is
still easy to pick up the stain.
12. ID Labs have a TdT antibody available, which needs only a high
temperature antigen unmasking for use in paraffin sections, with a
suggested dilution to 1:50 for an ABC system (this will be increased for
a more sensitive detection method).
13.Biocare currently have TdT.  We have a special antibody formula that
work well on the Ventana.
14.Zymed Laboratories, Inc. has polyclonal Rabbit anti-TdT for
immunohistostaining on formalin fixed paraffin-embedded sections.
15.I have used both Dako and Zymed Tdt antibodies on the Ventana system
and have
gotten great results with both ,but the zymed dilution was higher and a
retrieval was all I needed, I used them on the Ventana Es and the Nexes,
a 32 min incubation.

Thanks Again,
Dan Botsford
Windsor Regional Hospital
Windsor, Ontario, Canada.

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